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Detection of Outbreak-Associated Human Caliciviruses in Groundwater By Rt-Pcr

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Abstract:Human caliciviruses (HuCV) are a major worldwide cause of food and waterborne outbreaks of acute nonbacterial gastroenteritis, and have been placed on the U.S. Environmental Protection Agency's (U.S. EPA) Contaminant Candidate List (CCL) of agents to be considered for regulatory action. This activity has led to the development of a method to measure HuCV occurrence in drinking water. Viruses are considered the likely agent in over half of the waterborne outbreaks, but they have rarely been detected. HuCV are non-culturable and can only be detected in concentrated water samples using RT-PCR. In 2001, the U.S.EPA participated in the investigation of two HuCV outbreaks in the state of Wyoming. The first outbreak was recorded at a wintertime vacation lodge in northern Wyoming and the second during the summer at a restaurant in central Wyoming. For each outbrak a well water sample was concentrated for virus using a 1MDS positively charged cartridge filter. Filters were eluted with beef extract and the eluates concentrated with celite. The concentrated eluates were treated to remove PCR inihibitors and then assayed by RT-PCR using several HuCV-specific primer sets. HuCV were detected in the concentrated water samples from both outbreaks. Sequencing of the cloned RT-PCR products demonstrated the presence of a genogroup II, subtype 1 HuCV in the first outbreak and a genogroup II, subtype 3 HuCV in the second outbreak. These results demonstrate the utility of the RT-PCR method for investigations of waterborne outbreaks of suspected viral origin.
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Citation:Williantrue, S., S. Parshionikar, C. Newport, D. Robbins, and G. S. Fout. Detection of Outbreak-Associated Human Caliciviruses in Groundwater By Rt-Pcr. Presented at American Society for Virology Annual Meeting, Lexington, KY, July 20-24, 2002.
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Contact: Mary P. O'Bryant - (919)-541-4871 or obriant.mary@epa.gov
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Division: Microbiological & Chemical Exposure Assessment Division
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Branch: Biohazard Assessment Research Branch
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Product Type: Sympos/Conf
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Presented: 07/22/2002
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Related Entries:
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Bullet Item Measure Occurrence and Exposure to Waterborne Human Viruses
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Last Updated on Monday, October 22, 2007
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